Removal from adherent culture contributes to apoptosis in human bone marrow mesenchymal stem cells

Abstract

Mesenchymal stem cells (MSCs) are routinely isolated due to their adherence to tissue culture plates and their in vitro growth characteristics. Expansion of MSCs in adherent cultures is the only way to obtain sufficient cells for use in either clinical or research settings. MSCs have tremendous potential in myocardial repair treatment by cell therapy techniques, however, a large number of MSCs die from apoptosis following transplantation. Previous studies have examined the factors contributing to the survival of transplanted cells, but little is known about the effect of removal from adherent culture conditions on apoptosis of the MSCs. In the present study, human bone marrow MSCs were expanded in adherent cultures. Then apoptosis rates were examined at different time points in MSCs cultured in nonadherent conditions (ultra-low-adherence plates) compared with MSCs cultured in adherent conditions (standard tissue culture plates). Flow cytometry analysis suggested that cell apoptosis increased when MSCs were cultured in nonadherent culture conditions. In addition, western blot and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that caspase-3, −7 and −9 were involved in this process. The present study demonstrated that loss of culture adherence increases apoptosis of human MSCs. The present findings may provide new insight into the factors affecting MSC survival after transplantation.

Integration of Epigallocatechin Gallate in Gelatin Sponges Attenuates Matrix Metalloproteinase-Dependent Degradation and Increases Bone Formation

“…Indeed, in our immunohistological staining of actin using phalloidin, we found that cells were distributed along the residual gelatin of vhEGCG-GSs at 4 weeks, and sporadic cells were present at bone defect areas in the vhGS group. The attached cells, similar to osteoblasts and multipotent progenitor cells associated with bone formation, require robust cell attachment for survival [33,34]. Rapid digestion of gelatin by MMP-2 and -9 could yield a fragile, brittle scaffold for cell attachment and prevent bone formation by vhGSs, whereas vhEGCG-GSs seemed to provide a sufficient scaffold for cells via the inhibitory effects of MMPs.…”

Coupling normothermic machine perfusion with mesenchymal stem cell bioreactors: benefits exerted on the rat liver

“…We set up an innovative NMP perfusion system coupled with a MSCs-bioreactor with the aim to improve the application of cell-based therapies during NMP while avoiding any potential drawback related to stem cell direct administration 18,[32][33][34][35][36] . The particular con guration of our platform allowed a bidirectional communication between the liver and MSCs, enabling the cells to be primed with the perfusate enriched by the molecules secreted from the hepatic cells.…”

“…Stem cell cryopreservation itself can further reduce survival and trigger oxidative stress 32 . In addition, since MSCs are typically plastic-adherent in vitro, keeping these cells in suspension lead to apoptosis activation 33 . Direct cell administration and their migration into the graft raise other important concerns related to the risk of microvascular embolism 34,35 and unintended dedifferentiation of MSCs 36 .…”

Protective effects of α‑2‑macroglobulin on human bone marrow mesenchymal stem cells in radiation injury

“…In addition, ROS can initiate chain reactions, and react with a variety of unsaturated fatty acids and cholesterol on the cell membrane, resulting in oxidative damage to cells and eventually apoptosis. Cell death via apoptosis serves a role in maintaining normal tissue morphology and cell death, which are under the control of apoptosis-associated genes ( 31 , 32 ).…”